LOCUS       V034636                 9086 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034636
VERSION     V034636
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             775..3012
                     /label="EXT1(NM_000127)"
                     /note="EXT1(NM_000127)"
                     /gene="EXT1"
     CDS             3028..3084
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             3085..3795
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     misc_feature    3862..4450
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4516..4564
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4766..5194
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5208..5537
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5604..6398
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6574..6695
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6744..6760)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6768..6784
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6792..6822)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6837..6858
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7146..7734)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7905..8765)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8766..8870)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatgcag
      781 gccaaaaaac gctatttcat cctgctctca gctggctctt gtctcgccct tttgttttat
      841 ttcggaggct tgcagtttag ggcatcgagg agccacagcc ggagagaaga acacagcggt
      901 aggaatggct tgcaccaccc cagtccggat catttctggc cccgcttccc ggacgctctg
      961 cgccccttcg ttccttggga tcaattggaa aacgaggatt ccagcgtgca catttccccc
     1021 cggcagaagc gagatgccaa ctccagcatc tacaaaggca agaagtgccg catggagtcc
     1081 tgcttcgatt tcaccctttg caagaaaaac ggcttcaaag tctacgtata cccacagcaa
     1141 aaaggggaga aaatcgccga aagttaccaa aacattctag cggccatcga gggctccagg
     1201 ttctacacct cggaccccag ccaggcgtgc ctctttgtcc tgagtctgga tactttagac
     1261 agagaccagt tgtcacctca gtatgtgcac aatttgagat ccaaagtgca gagtctccac
     1321 ttgtggaaca atggtaggaa tcatttaatt tttaatttat attccggcac ttggcctgac
     1381 tacaccgagg acgtggggtt tgacatcggc caggcgatgc tggccaaagc cagcatcagt
     1441 actgaaaact tccgacccaa ctttgatgtt tctattcccc tcttttctaa ggatcatccc
     1501 aggacaggag gggagagggg gtttttgaag ttcaacacca tccctcctct caggaagtac
     1561 atgctggtat tcaaggggaa gaggtacctg acagggatag gatcagacac caggaatgcc
     1621 ttatatcacg tccataacgg ggaggacgtt gtgctcctca ccacctgcaa gcatggcaaa
     1681 gactggcaaa agcacaagga ttctcgctgt gacagagaca acaccgagta tgagaagtat
     1741 gattatcggg aaatgctgca caatgccact ttctgtctgg ttcctcgtgg tcgcaggctt
     1801 gggtccttca gattcctgga ggctttgcag gctgcctgcg tccctgtgat gctcagcaat
     1861 ggatgggagt tgccattctc tgaagtgatt aattggaacc aagctgccgt cataggcgat
     1921 gagagattgt tattacagat tccttctaca atcaggtcta ttcatcagga taaaatccta
     1981 gcacttagac agcagacaca attcttgtgg gaggcttatt tttcttcagt tgagaagatt
     2041 gtattaacta cactagagat tattcaggac agaatattca agcacatatc acgtaacagt
     2101 ttaatatgga acaaacatcc tggaggattg ttcgtactac cacagtattc atcttatctg
     2161 ggagattttc cttactacta tgctaattta ggtttaaagc ccccctccaa attcactgca
     2221 gtcatccatg cggtgacccc cctggtctct cagtcccagc cagtgttgaa gcttctcgtg
     2281 gctgcagcca agtcccagta ctgtgcccag atcatagttc tatggaattg tgacaagccc
     2341 ctaccagcca aacaccgctg gcctgccact gctgtgcctg tcgtcgtcat tgaaggagag
     2401 agcaaggtta tgagcagccg ttttctgccc tacgacaaca tcatcacaga cgccgtgctc
     2461 agccttgacg aggacacggt gctttcaaca acagaggtgg atttcgcctt cacagtgtgg
     2521 cagagcttcc ctgagaggat tgtggggtac cccgcgcgca gccacttctg ggataactct
     2581 aaggagcggt ggggatacac atcaaagtgg acgaacgact actccatggt gttgacagga
     2641 gctgctattt accacaaata ttatcactac ctatactccc attacctgcc agccagcctg
     2701 aagaacatgg tggaccaatt ggccaattgt gaggacattc tcatgaactt cctggtgtct
     2761 gctgtgacaa aattgcctcc aatcaaagtg acccagaaga agcagtataa ggagacaatg
     2821 atgggacaga cttctcgggc ttcccgttgg gctgaccctg accactttgc ccagcgacag
     2881 agctgcatga atacgtttgc cagctggttt ggctacatgc cgctgatcca ctctcagatg
     2941 aggctcgacc ccgtcctctt taaagaccag gtctctattt tgaggaagaa ataccgagac
     3001 attgagcgac ttgaattcgg aagcggagct actaacttca gcctgctgaa gcaggctggc
     3061 gacgtggagg agaaccctgg acctatggtg agcaagggcg aggaggataa catggccatc
     3121 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     3181 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     3241 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     3301 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     3361 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     3421 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     3481 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     3541 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     3601 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     3661 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     3721 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     3781 gagctgtaca agtaactcga gtctagaggg ccccttaagg gttcgatccc taccggttag
     3841 taatgagttt gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact
     3901 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3961 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4021 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4081 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4141 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4201 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     4261 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     4321 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4381 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4441 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4501 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     4561 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4621 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4681 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     4741 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4801 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4861 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4921 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4981 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5041 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5101 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     5161 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     5221 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5281 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5341 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5401 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5461 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5521 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5581 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5641 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5701 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5761 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5821 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5881 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5941 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     6001 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     6061 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     6121 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6181 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6241 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6301 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6361 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     6421 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     6481 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6541 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     6601 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6661 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     6721 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6781 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6841 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6901 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6961 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     7021 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     7081 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     7141 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7201 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7261 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7321 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7381 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7441 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7501 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7561 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     7621 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7681 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7741 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7801 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7861 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7921 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7981 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     8041 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     8101 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     8161 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     8221 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     8281 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8341 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8401 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     8461 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8521 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8581 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8641 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8701 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8761 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8821 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8881 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc
     8941 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     9001 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     9061 acaattgcat gaagaatctg cttagg
//
